Antibiotic resistance is a pressing issue that is widely covered in college biology courses. Here, we present a hands-on laboratory exercise to experimentally demonstrate how antibiotics work as well as how cells can become resistant to antibiotics. The use of freeze-dried, cell-free protein synthesis reactions instead of live bacterial cultures makes this exercise highly accessible with minimal equipment requirements. Students add antibiotics that inhibit protein production to cell-free reactions and observe the effects using a fluorescent protein as a visual readout of translation. Additionally, students express an antibiotic resistance gene to test how resistance genes can rescue translation inhibition in an antibiotic-specific manner. The initial implementation of this exercise demonstrated that the activity is easy to implement, engages student interest in topics related to antibiotic resistance, and has the potential to be widely integrated into undergraduate courses.
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